Aβ1−42 was bought from Sigma-Aldrich (St. Louis, USA) and GL Biochem. (Beijing, China) as the lyophilized powder. Hexafluoroisopropanol (HFIP) was purchased from Merck Co. (Darmstadt, Germany). Thioflavin T (ThT), thiazolyl blue tetrazolium bromide (MTT), 8-Anilinonaphthalene-1-sulfonate (ANS), and Hoechst were purchased from Sigma-Aldrich Chem. Co. (St. Louis, USA). The PC12 rat pheochromocytoma cell line was purchased from Pasture Institute (Tehran, Iran). Cell culture plates were acquired from SPL (Beijing, China). Primary antibodies (ab201060 and ab224275) and the secondary antibody (ab6721) were bought from Abcam (Abcam Inc., Cambridge, MA). ECL Plus Kit was purchased from Bio-Rad (Bio-Rad Laboratories Inc., Hercules, CA, USA). Alexa Fluor 594 (clone Poly4064) was purchased from BioLegend (San Diego, CA, USA). PVDF was purchased from GE Healthcare (Biosciences, Stockholm, Sweden). All other materials were of analytical grade.
Ab224275
Manufactured by Abcam
Sourced in United States
Ab224275 is a recombinant protein that can be used as a positive control in western blot experiments. It is a full-length human protein expressed in and purified from HEK293 cells.
Automatically generated - may contain errors
Lab products found in correlation
Thiazolyl blue tetrazolium bromide mtt, by Merck Group (1 mentions)
Cell culture plate, by SPL Life Sciences (1 mentions)
Hoechst, by Merck Group (1 mentions)
Aβ1 42, by Merck Group (1 mentions)
Polyvinylidene fluoride (pvdf), by GE Healthcare (1 mentions)
Thioflavin t tht, by Merck Group (1 mentions)
Ab6721, by Abcam (1 mentions)
Ab201060, by Abcam (1 mentions)
Ecl plus kit, by Bio-Rad (1 mentions)
Acetonitrile acn, by Merck Group (1 mentions)
Phosphate buffered saline tablets pbs ph 7, by Merck Group (1 mentions)
Amyloid beta 1 42, by Abcam (1 mentions)
Vibratome, by Leica (1 mentions)
Dapi solution, by Solarbio (1 mentions)
4 protocols using ab224275
1
Amyloid-β Peptide Aggregation Assay
2
Electrochemical Detection of Amyloid-Beta
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Materials 4-nitrobenzenediazonium tetrafluoroborate 97% (4-NPD), tetrabutylammonium tetrafluoroborate 99% (BU4NBF4), 1-ethyl-3-(3-(dimethylamino)-propyl) carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), ferrocene carboxylic acid (FCA), potassium chloride, acetonitrile (ACN) and phosphate-buffered saline tablets (PBS; pH 7.4) were all purchased from Sigma Aldrich (St. Louis, MO, USA) and used as received. All reagents were of analytical grade, and the deionised water was used for the cleaning of the glassy carbon electrodes. Highly ordered pyrolytic graphite (HOPG) was purchased from MikroMasc (Wetzlar, Germany) with the following specifications: grade ZYA, mosaic spread with the value of 0.4° ± 0.1°, double-sided and with thickness 1 mm and the size of 10 × 10 mm2.
Biomarker amyloid-beta peptides Aβ ((1-42) (ab120301)) and anti-beta amyloid antibody (ab224275), specific to Aβ (1-42) peptide Aβ, were purchased from Abcam (Cambridge, UK).
Amyloid-beta (1-42) was supplied in the lyophilised form by Abcam (Cambridge, UK). It was then dissolved in 10 mM sodium hydroxide, followed by gentle Vortex for less than 1 min to make a homogeneous solution. The peptide stock solution was then aliquoted and stored at −20 °C. Under these highly alkaline conditions, the peptide is fully dissolved and exists only as monomers [43 (link)].
Biomarker amyloid-beta peptides Aβ ((1-42) (ab120301)) and anti-beta amyloid antibody (ab224275), specific to Aβ (1-42) peptide Aβ, were purchased from Abcam (Cambridge, UK).
Amyloid-beta (1-42) was supplied in the lyophilised form by Abcam (Cambridge, UK). It was then dissolved in 10 mM sodium hydroxide, followed by gentle Vortex for less than 1 min to make a homogeneous solution. The peptide stock solution was then aliquoted and stored at −20 °C. Under these highly alkaline conditions, the peptide is fully dissolved and exists only as monomers [43 (link)].
3
Amyloid-Beta Immunohistochemistry in Murine Hippocampi
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The tissues of the mice hippocampi xed by 4% paraformaldehyde were embedded in para n and sliced coronally into 5-μm sections using a vibratome (Leica, Germany) . The sections were performed for antigen repair at 99.9 ° C for 30 min after depara nation and then incubated for 10 min in 3% H2O2, and incubated with primary rabbit polyclonal antibodies against Aβ 1-40 (bs-0877R, 1:100, Boiss, China), primary Rabbit monoclonal antibodies against Aβ 1-42 (ab224275, 1:200, Abcam, Cambridge, MA, USA) overnight at 4 °C. Then the sections were incubated with biotinlabeled secondary antibodies (1:300, 865002, R&D Systems) for 30 min at 37 °C and the Cell and Tissue Staining Kit (CTS005, Anti-Rabbit HRP-DAB System, R&D Systems) was used to detect the positive staining area. The images were captured by a camera (Nikon, 90i, Tokyo, Japan). And 3 sections per mouse were being counted in a blind manner.
4
Quantifying Amyloid-Beta Deposition
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N2a neuronal cells xed with 4%paraformaldehyde were permeated by 0.5% TritonX-100 in PBS, and then blocked in 2% BSA at 37•C for 1 h and then incubated with β-Amyloid (1-40 Speci c) (D8Q7I) Rabbit mAb (12990, 1 : 600, Cell Signaling Technology, USA), Rabbit monoclonal antibodies against Aβ 1-42 (ab224275, 1: 200, Abcam, Cambridge, MA, USA) for 1 day at 4•C. After washing with PBS, the cells were incubated with Goat Anti-Rabbit IgG H&L FITC L30113, 1:200; Signalway Antibody, USA at room temperature for 1h in the dark. After washing with PBS, the sections were incubated with DAPI solution (Solarbio, Beijing, China) at room temperature for 5min and mounted in the Vectashield anti-fading medium (Solarbio, Beijing, China). Fluorescent images were captured by LEICA DMi8 and quanti ed by the ImageJ software (1.41V, US National Institutes of Health).
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